Susanne Polywka

Reactivation of Hepatitis B Virus Replication Accompanied by Acute Hepatitis in Patients Receiving Highly Active Antiretroviral Therapy

We describe 2 patients who were initially positive for antibodies to hepatitis B surface antigen and who experienced a strong and sudden increase of hepatitis B virus (HBV) replication during highly active antiretroviral therapy (HAART). We found that reactivation of HBV replication during HAART can occur independently of lamivudine resistance or withdrawal of lamivudine, and in spite of increasing CD4(+) cell counts.

Low Risk of Vertical Transmission of Hepatitis C Virus by Breast Milk

To evaluate the risk of hepatitis C virus (HCV) transmission via breast milk, we collected 76 samples of breast milk from 73 chronically HCV-infected women and serum samples from their 76 perinatally HCV-exposed children. Enzyme immunoassay and strip immunoblot assay were used for detection of antibodies to HCV, and reverse transcriptase-polymerase chain reaction analysis was used for detection of HCV RNA. None of the 76 samples of breast milk contained HCV RNA, whereas 37 (59.7%) of 62 mothers tested for HCV RNA had HCV viremia. Only 1 of the 76 breast-fed infants had evidence of HCV infection. Because HCV infection in this child was detected 1 month after birth, it seems unlikely that it was transmitted by breast-feeding. These results indicate that HCV infection in pregnant women should not be a contra-indication for breast-feeding.

Guidelines for the Treatment of Hepatitis C Virus Infection in Injection Drug Users: Status Quo in the European Union Countries

Treatment guidelines are considered to be an important tool in steering patients to medical treatment. This study was conducted to analyze guidelines for the treatment of hepatitis C virus (HCV) infection in injection drug users (IDUs) in the European Union (EU) countries as a component of treatment access. National and international databases, expert contacts, professional societies, and health administrations were approached to acquire guidelines. According to their quality standard, guidelines were divided into expert opinions, semiofficial guidelines, official guidelines, and consensus processes. Recommendations for the treatment of HCV infection in IDUs vary substantially, from lack of recommendations and outright treatment disapproval to recommendations for treatment under specified circumstances. Recent guidelines that apply qualified process procedures that include literature research tend to be more permissive. Qualified guideline processes in each EU country and subsequently renewed pan-European guidelines are needed.

Application of nested PCR and mass spectrometry for DNA-based virus detection: HBV-DNA detected in the majority of isolated anti-HBc positive sera

DNA preparations from three different groups of serum samples were examined for HBV-DNA via a nested polymerase chain reaction assay (lower detection limit: 10 viral genomes in 100 microliters serum): Group I consisted of 11 uninfected control sera, group II consisted of sera obtained from 11 HBV infected patients and group III consisted of 21 isolated anti-HBc positive samples. The 21 samples from group III were HBV-DNA negative according to a conventional non-nested PCR assay and hybridization with a 32P-labelled probe. Using nested PCR and mass spectrometry, HBV-DNA was detected in none of group I and in all of group II samples. In 11 out of 21 (52%) of the isolated anti-HBc positive sera from group III, HBV-DNA was detected. No correlation was observed between HBV-DNA positivity and anti-HBc titers. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided a fast, sensitive and non-radioactive assay for the detection of PCR products without the need for gel electrophoresis or hybridization with labelled probes.

Human liver chimeric mice as a new model of chronic hepatitis E virus infection and preclinical drug evaluation

BACKGROUND & AIMS Hepatitis E virus (HEV) is a major cause of acute hepatitis as well as chronic infection in immunocompromised individuals; however, in vivo infection models are limited. The aim of this study was to establish a small animal model to improve our understanding of HEV replication mechanisms and permit the development of effective therapeutics. METHODS UPA/SCID/beige mice repopulated with primary human hepatocytes were used for infection experiments with HEV genotype (GT) 1 and 3. Virological parameters were determined at the serological and intrahepatic level by real time PCR, immunohistochemistry and RNA in situ hybridization. RESULTS Establishment of HEV infection was achieved after intravenous injection of stool-derived virions and following co-housing with HEV-infected animals but not via inoculation of serum-derived HEV. GT 1 infection resulted in a rapid rise of viremia and high stable titres in serum, liver, bile and faeces of infected mice for more than 25 weeks. In contrast, viremia in GT 3 infected mice developed more slowly and displayed lower titres in all analysed tissues as compared to GT 1. HEV-infected human hepatocytes could be visualized using HEV ORF2 and ORF3 specific antibodies and HEV RNA in situ hybridization probes. Finally, six-week administration of ribavirin led to a strong reduction of viral replication in the serum and liver of GT 1 infected mice. CONCLUSION We established an efficient model of HEV infection to test the efficacy of antiviral agents and to exploit mechanisms of HEV replication and interaction with human hepatocytes in vivo.

Persistent hepatitis D virus mono-infection in humanized mice is efficiently converted by hepatitis B virus to a productive co-infection

BACKGROUND & AIMS Clinical studies have shown that hepatitis delta virus (HDV) infection can persist for years and intrahepatic latency of the large delta antigen (HDAg) has been detected following liver transplantation. However, large HDAg arising via RNA-editing is associated with increasing amounts of non-infectious HDV quasi-species. This study investigated whether HDV could persist intrahepatically in the absence of HBV in vivo and whether infectious HDV could subsequently be released following HBV super-infection. METHODS Humanized mice were infected with HDV particles lacking HBV. To test for rescue of latent HDV infection 3 and 6 weeks HDV mono-infected mice were super-infected with HBV. Viral loads and cell toxicity were determined by qRT-PCR and immunohistochemistry. RESULTS The presence of HDAg-positive human hepatocytes determined after 2, 3, and 6 weeks of HDV inoculation demonstrated establishment and maintenance of intrahepatic HDV mono-infection. Although intrahepatic amounts of large HDAg and edited HDV RNA forms increased over time in HDV mono-infected livers, HBV super-infection led to prompt viremia development (up to 10(8) HDV RNA and 10(7) HBV-DNA copies/ml) even after 6 weeks of latent mono-infection. Concurrently, the number of HDAg-positive human hepatocytes increased, demonstrating intrahepatic HDV spreading. The infectivity of the rescued HDV virions was verified by serial passage in naive chimeric mice. CONCLUSIONS HDV mono-infection can persist intrahepatically for at least 6 weeks before being rescued by HBV. Conversion of a latent HDV infection to a productive HBV/HDV co-infection may contribute to HDV persistence even in patients with low HBV replication and in the setting of liver transplantation.

Efficacy and safety of sofosbuvir/ledipasvir for the treatment of patients with hepatitis C virus re-infection after liver transplantation.

Hepatitis C virus (HCV) infection is associated with a particularly poor outcome after liver transplantation. In December 2014, sofosbuvir/ledipasvir (SOF/LDV) fixed‐dose combination (FDC) was approved for HCV genotype 1 and 4 in Europe. In orthotopic liver transplantation (OLT) recipients, the interferon‐free treatment of HCV re‐infection with novel direct‐acting antivirals has been demonstrated to be safe and effective in clinical trials, but real‐world data are missing. The aim of this study was to investigate the safety and efficacy of SOF/LDV FDC in OLT recipients in the real‐life setting.

Vertebral osteomyelitis due to Rhodococcus equi in a liver transplant recipient

Rhodococcus equi is a rare but well-documented cause of cavitary pneumonia in immunocompromised patients. In this report the first case of R. equi infection manifesting as vertebral osteomyelitis is described. A 39-year-old liver transplant recipient presented with recurrent pneumonia and a pleura-based lung abscess and subsequently developed osteomyelitis of the lower thoracic spine. Surgical debridement and prolonged treatment with rifabutin and clarithromycin resulted in clinical cure. In the literature, 12 other cases of R. equi infection in solid-organ transplant recipients have been reported. Ten of these patients had documented pulmonary disease and seven had extrapulmonary manifestations. Prolonged antibiotic therapy and surgical drainage resulted in clinical improvement in > 90% of the reported cases.

Greater Amount of HCV-RNA in Tears Compared to Blood

Plasma, tear fluid and swabs from eye, nose and pharynx of 33 patients were examined for presence of hepatitis C virus (HCV) RNA by polymerase chain reaction (PCR). All samples from plasma, tear fluid and eyeswabs were found to show a positive reaction in HCV‐RNA PCR. Remarkably, we regularly found greater amounts of amplification products in tear fluid and eyeswabs compared to plasma using the same conditions for sample preparation.

Prolonged time until seroconversion among hemodialysis patients: the need for HCV PCR.

Objective: Patients on maintenance hemodialysis are known to have an elevated risk of acquiring hepatitis C virus (HCV) infection. Therefore, a reliable diagnosis of HCV infection is essential in order to prevent the spread of the disease in dialysis units. However, whether PCR examination is dispensable in hemodialysis patients has been debated. Methods: From 1995 to 2002, serum samples from all hemodialysis patients at our hospital (n = 1,774) were screened by serological assays and by polymerase chain reaction (PCR). Results: In 25 of these patients acute HCV infection was observed and in 11 patients HCV seroconversion was delayed for 3–16 months. During this time the infection was exclusively detectable by PCR. Conclusion: Despite the growing demand for cost-effectiveness in the health system, HCV PCR examination must remain an essential part of the routine screening in hemodialysis patients.